Practical High-performance Liquid Chromatography, 4th Edition

by Veronika R. Meyer (EMPA, St Gallen, Switzerland)

• ISBN13: 9780470093788
• ISBN10: 0470093781
• Edition: 4th
• Format: Paperback
• Copyright: 2004-10-01
• Publisher: Wiley-Interscience

Summary

Veronika Meyer's book on HPLC is a classic text and remains one of the few titles available on general HPLC. Following on from the success of the previous three editions, this new, fourth edition continues to provide users of HPLC in industry, government, and service laboratories, as well as postgraduate students, with a unified approach to HPLC and an equal treatment of the theory and practice of this important technique.The contents of this edition have been revised, expanded and updated. Where available, old literature references have been replaced by recent ones. New sections on the following topics have been included:Shelf-live of mobile phasesThe mixing crossPhase systems in ion chromatographyMeasurement uncertainty

Table of Contents

From the Preface to the First Edition	xiii
Preface to the Fourth Edition	xv
Important and Useful Equations for HPLC	1	(158)
1 Introduction	4	(10)
1.1 HPLC: A powerful separation method	4	(1)
1.2 A first HPLC experiment	4	(3)
1.3 Liquid chromatographic separation modes	7	(1)
1.4 The HPLC instrument	8	(1)
1.5 Safety in the HPLC laboratory	9	(1)
1.6 Comparison between high-performance liquid chromatography and gas chromatography	10	(1)
1.7 Pressure units	11	(1)
1.8 Length units	11	(1)
1.9 Scientific journals	12	(1)
1.10 Recommended books	12	(2)
2 Theoretical Principles	14	(38)
2.1 The chromatographic process	14	(2)
2.2. Band broadening	16	(4)
2.3 The chromatogram and its purport	20	(7)
2.4 Graphical representation of peak pairs with different degrees of resolution	27	(5)
2.5 Factors affecting resolution	32	(5)
2.6 Extra-column volumes (dead volumes)	37	(1)
2.7 Tailing	37	(5)
2.8 Peak capacity and statistical resolution probability	42	(3)
2.9 Effects of temperature in HPLC	45	(2)
2.10 The limits of HPLC	47	(5)
3 Pumps	52	(6)
3.1 General requirements	52	(1)
3.2 The short-stroke piston pump	52	(3)
3.3 Maintenance and repair	55	(1)
3.4 Other pump designs	56	(2)
4 Preparation of Equipment up to Sample Injection	58	(15)
4.1 Selection of mobile phase	58	(2)
4.2 Preparation of the mobile phase	60	(1)
4.3 Gradient systems	61	(2)
4.4 Capillary tubing	63	(3)
4.5 Fittings	66	(1)
4.6 Sample injectors	67	(4)
4.7 Sample solution and sample volume	71	(2)
5 Solvent Properties	73	(9)
5.1 Table of organic solvents	73	(2)
5.2 Solvent selectivity	75	(1)
5.3 Miscibility	76	(1)
5.4 Buffers	76	(3)
5.5 Shelf-life of mobile phases	79	(1)
5.6 The mixing cross	79	(3)
6 Detectors	82	(24)
6.1 General	82	(5)
6.2 UV detectors	87	(3)
6.3 Refractive index detectors	90	(2)
6.4 Fluorescence detectors	92	(1)
6.5 Electrochemical (amperometric) detectors	92	(2)
6.6 Light scattering detectors	94	(2)
6.7 Other detectors	96	(1)
6.8 Multiple detection	97	(1)
6.9 Indirect detection	98	(1)
6.10 Coupling with spectroscopy	99	(7)
7 Columns and Stationary Phases	106	(24)
7.1 Columns for HPLC	106	(2)
7.2 Precolumns	108	(1)
7.3 General properties of column packings	109	(5)
7.4 Silica	114	(1)
7.5 Chemically modified silica	115	(3)
7.6 Styrene-divinylbenzene	118	(4)
7.7 Some other stationary phases	122	(4)
7.8 Column care and regeneration	126	(4)
8 HPLC Column Tests	130	(16)
8.1 Simple tests for HPLC columns	130	(2)
8.2 Determination of particle size	132	(1)
8.3 Determination of breakthrough time	133	(2)
8.4 The test mixture	135	(3)
8.5 Dimensionless parameters for HPLC column characterization	138	(3)
8.6 The van Deemter equation from reduced parameters and its use in column diagnosis	141	(2)
8.7 Diffusion coefficients	143	(3)
9 Adsorption Chromatography	146	(13)
9.1 What is adsorption?	146	(3)
9.2 The eluotropic series	149	(3)
9.3 Selectivity properties of the mobile phase	152	(1)
9.4 Choice and optimization of the mobile phase	153	(1)
9.5 Applications	154	(5)
10 Reversed-Phase Chromatography	159	(19)
10.1 Principle	159	(2)
10.2 Mobile phases in reversed-phase chromatography	161	(2)
10.3 Solvent selectivity and strength	163	(4)
10.4 Stationary phases	167	(3)
10.5 Method development in reversed-phase chromatography	170	(3)
10.6 Applications	173	(2)
10.7 Hydrophobic interaction chromatography	175	(3)
11 Chromatography with Chemically Bonded Phases	178	(5)
11.1 Introduction	178	(1)
11.2 Properties of some stationary phases	178	(5)
12 Ion-Exchange Chromatography	183	(12)
12.1 Introduction	183	(1)
12.2 Principle	183	(1)
12.3 Properties of ion exchangers	184	(2)
12.4 Influence of the mobile phase	186	(2)
12.5 Special possibilities of ion exchange	188	(2)
12.6 Practical hints	190	(2)
12.7 Applications	192	(3)
13 Ion-Pair Chromatography	195	(7)
13.1 Introduction	195	(1)
13.2 Ion-pair chromatography in practice	196	(2)
13.3 Applications	198	(1)
13.4 Appendix: UV detection using ion-pair reagents	199	(3)
14 Ion Chromatography	202	(5)
14.1 Principle	202	(1)
14.2 Suppression techniques	203	(1)
14.3 Phase systems	203	(3)
14.4 Applications	206	(1)
15 Size-Exclusion Chromatography	207	(15)
15.1 Principle	207	(3)
15.2 The calibration chromatogram	210	(3)
15.3 Molecular mass determination by means of size-exclusion chromatography	213	(2)
15.4 Coupled size-exclusion columns	215	(2)
15.5 Phase systems	217	(1)
15.6 Applications	218	(4)
16 Affinity Chromatography	222	(6)
16.1 Principle	222	(1)
16.2 Affinity chromatography as a special case of HPLC	223	(2)
16.3 Applications	225	(3)
17 Choice of Method	228	(7)
18 Solving the Elution Problem	235	(20)
18.1 The elution problem	235	(1)
18.2 Solvent gradients	235	(6)
18.3 Column switching	241	(3)
18.4 Optimization of an isocratic chromatogram using four solvents	244	(3)
18.5 Optimization of the other parameters	247	(6)
18.6 Mixed stationary phases	253	(2)
19 Analytical HPLC	255	(30)
19.1 Qualitative analysis	255	(2)
19.2 Trace analysis	257	(4)
19.3 Quantitative analysis	261	(5)
19.4 Recovery	266	(2)
19.5 Peak-height and peak-area determination for quantitative analysis	268	(4)
19.6 Integration errors	272	(2)
19.7 The detection wavelength	274	(1)
19.8 Apparatus test, validation and system suitability test	275	(3)
19.9 Measurement uncertainty	278	(1)
19.10 Derivatization	279	(3)
19.11 Unexpected peaks: ghost and system peaks	282	(3)
20 Preparative HPLC	285	(12)
20.1 Problem	285	(1)
20.2 Preparative HPLC in practice	286	(3)
20.3 Overloading effects	289	(3)
20.4 Fraction collection	292	(2)
20.5 Recycling	294	(1)
20.6 Displacement chromatography	294	(3)
21 Separation of Enantiomers	297	(14)
21.1 Introduction	297	(2)
21.2 Chiral mobile phases	299	(1)
21.3 Chiral liquid stationary phases	300	(1)
21.4 Chiral solid stationary phases	301	(7)
21.5 Indirect separation of enantiomers	308	(3)
22 Special Possibilities	311	(9)
22.1 Micro and capillary HPLC	311	(2)
22.2 High-speed and super-speed HPLC	313	(3)
22.3 HPLC with supercritical mobile phases	316	(2)
22.4 Electrochromatography	318	(2)
23 Appendix 1: Applied HPLC Theory	320	(10)
24 Appendix 2: How to Perform the Instrument Test	330	(7)
24.1 Introduction	330	(1)
24.2 The test procedure	331	(4)
24.3 Documentation, limiting values and tolerances	335	(2)
25 Appendix 3: Troubleshooting	337	(8)
26 Appendix 4: Column Packing	345	(4)
Index of Separations	349	(2)
Index	351